Selection of resistance breaking strains of tomato spotted wilt tospovirus

In glasshouse tests, sap from plants infected with 15 different isolates of tomato spotted wilt tospovirus (TSWV) from three Australian states was inoculated to nine genotypes of tomato carrying TSWV resistance gene Sw-5 or one of its alleles. A further two resistant tomato genotypes were inoculated with four isolates each. The normal response in resistant genotypes was development of necrotic local lesions in inoculated leaves without systemic invasion, but 22/752 plants also developed systemic reactions in addition to local hypersensitive ones. Using cultures from two of these systemically infected plants and following four cycles of subculture in TSWV resistant tomato plants, two isolates were obtained that gave susceptible type systemic reactions but no necrotic spots in inoculated leaves of resistant tomatoes. When these two isolates, Da_WA-1d and To_TAS-1d, were maintained by repeated subculture for 10 successive cycles in Nicotiana glutinosa or a susceptible tomato genotype, they still induced susceptible type systemic reactions when inoculated to resistant tomato plants. They were therefore stable resistance breaking isolates as regards overcoming gene Sw-5. When resistance-breaking isolate Da_WA^-1ld multiplied together with original isolate Da_WA-l in susceptible tomato, it was fully competitive with the original isolate. However, when Da_WA-ld and To_TAS-ld were inoculated to TSWV resistant Lycopersicon peruvianum lines PI 128660R and PI 128660S and to TSWV resistant Capsicum chinense lines PI 152225, PI 159236 and AVRDC CO0943, they failed to overcome the resistance, producing only necrotic local lesions without systemic infection. Thus, although the ease of selection, stability and competitive ability of resistance breaking isolates of TSWV is cause for concern, L. peruvianum and C. chinense lines are available which are effective against them. The effectiveness of the resistance to TSWV in nine tomato genotypes was examined in a field experiment. Spread was substantial in the susceptible control genotype infecting 42% of plants. Resistance was ineffective in cv. Bronze Rebel, 26% of plants developing infection. In contrast, it held up well in the other eight resistant genotypes with only 1–3 or no plants of each becoming infected. Accumulated numbers of Thrips tabaci, Frankliniella occidentalis and F. schultzei were closely correlated with TSWV spread.     

A Comparison of the Disease Reactions of Stems and Detached Leaves of Soil and in vitro Grown Plants and Regenerants of Oilseed Rape to Leptosphaeria maculans and Protocols for Selection for Novel Disease Resistance

Protocols for selecting plant tissues of winter oilseed rape (Brassica napus subsp. oleifera) with resistance to Leptosphaeria maculans by either stem or leaf inoculation of both soil and in vitro grown plant material are described. The stem inoculation procedure gave good correlation (r = 0. 92) between the 50 day stem disease scores of eight out of nine cultivars of soil grown winter oilseed rape inoculated with isolate 41A4 of L. maculans and the N. A. B. esistance ratings or resistance data from field trials. The exception was the cultivar Liradonna. Inoculation of stems of five cultivars with isolates 41A4, 433 and 478 indicated a range of isolate virulence 478 > 41A4 > 433. This was the inverse of that observed in leaf inoculations. Application of the stem inoculation procedure to in vitro shoot cultures allowed differentiation of resistant and susceptible cultivars, including the cultivar Liradonna, after 20 days incubation at 20°C. The protocol was also applicable to plantlets regenerated from thin cell layer explants grown in vitro. Inoculations with isolate 433 allowed the differentiation of resistant, intermediately resistant and susceptible leaf material of soil grown plants, when leaf discs from young leaves were incubated on water agar supplemented with BAP (1 × 10^?5 M) at 25°C for 10 days. Intermediately resistant leaves were resistant after 10 days and susceptible after 15 days of incubation. Leaves of shoot cultures grown in vitro were more susceptible than the corresponding soil grown material. However, inoculation of old leaves with isolate 41A4 (an isolate of less virulence on leaves than 433) distinguished the cultivars after 15 days of incubation. These protocols allow the accurate assessment of resistance to L. maculans at the stem or leaf level and are of use in traditional as well as in vitro selection programmes.     

Effects of Meloidogyne spp. and Rhizoctonia solani on the Growth of Grapevine Rootings

A disease complex involving Meloidogyne incognita and Rhizoctonia solani was associated with stunting of grapevines in a field nursery. Nematode reproduction was occurring on both susceptible and resistant cultivars, and pot experiments were conducted to determine the virulence of this M. incognita population, and of M. javanica and M. hapla populations, to V. vinifera cv. Colombard (susceptible) and to V. champinii cv. Ramsey (regarded locally as highly resistant). The virulence of R. solani isolates obtained from roots of diseased grapevines also was determined both alone and in combination with M. incognita. Ramsey was susceptible to M. incognita (reproduction ratio 9.8 to 18.4 in a shadehouse and heated glasshouse, respectively) but was resistant to M. javanica and M. hapla. Colombard was susceptible to M. incognita (reproduction ratio 24.3 and 41.3, respectively) and M. javanica. Shoot growth was suppressed (by 35%) by M. incognita and, to a lesser extent, by M. hapla. Colombard roots were more severely galled than Ramsey roots by all three species, and nematode reproduction was higher on Colombard. Isolates of R. solani assigned to putative anastomosis groups 2-1 and 4, and an unidentified isolate, colonized and induced rotting of grapevine roots. Ramsey was more susceptible to root rotting than Colombard. Shoot growth was inhibited by up to 15% by several AG 4 isolates and by 20% by the AG 2-1 isolate. AG 4 isolates varied in their virulence. Root rotting was higher when grapevines were inoculated with both M. incognita and R. solani and was highest when nematode inoculation preceded the fungus. Shoot weights were lower when vines were inoculated with the nematode 13 days before the fungus compared with inoculation with both the nematode and the fungus on the same day. It was concluded that both the M. incognita population and some R. solani isolates were virulent against both Colombard and Ramsey, and that measures to prevent spread in nursery stock were therefore important.     

Host specificity,pathogenicity and the presence of virulence genes in Iranian strains of Pseudomonas syringae pv. syringae from different hosts

Pseudomonas syringae pv. syringae (Pss) strains were isolated from almond, apricot, peach, pear, sweet cheery and wheat in Kohgiluye and Boyer-Ahmad, Kordestan, Fras and Chaharmahal and Bakhtiari provinces of Iran. The strains were examined for host specificity, the presence of virulence genes and pathogenicity on different hosts. After inoculation of isolates, in compatible reactions bacterial populations increased within six days of inoculation and final cell numbers increased several-fold over initial inoculum levels, but in incompatible reactions, bacterial populations declined within four days of inoculation. Almond, sweet cherry and wheat isolates induced progressive necrotic symptoms on almond leaves and stems. Apricot, peach and sweet cherry isolates induced necrotic lesions when inoculated on apricot leaves. On pear leaves and stems, only the pear isolate incited pathogenic reaction and isolates from other hosts did not. The syrB gene was detected in all of the tested isolates. Almond and pear isolates did not have the syrD gene. The sypA gene was detected in the almond, peach, pear and sweet cherry isolates while the sypB gene was detected in the apricot, peach, sweet cherry and wheat isolates. Almond, apricot, pear and wheat isolates gave negative results for the detection of nit gene. The gene Ach, was detected only in the peach isolate and gene hrmA, was detected only in the wheat isolate. This study indicates that host specificity exists among different Pss strains, and genes responsible for syringomycin and syringopeptin production contribute to the virulence of Pss strains.     

Interactions of Puccinia hordei and Erysiphe graminis on seedling barley

The development of Puccinia hordei on the first leaf of barley seedlings previously inoculated with Erysiphe graminis was compared with that on uninoculated leaves of comparable age. On cv. Zephyr, more rust pustules developed when leaves were inoculated with both fungi within 24 h but fewer pustules if the period between the two inoculations was longer than 2 days. The reduction in numbers of rust pustules was especially marked where leaves were previously inoculated with many conidia of E. graminis. The size of rust pustules was reduced whatever the period between the two inoculations. Arresting mildew development by applying ethirimol as a soil drench to pots of seedlings inoculated with E. graminis 6 days previously, or floating segments of leaves inoculated with both fungi on 2% sucrose, in part counteracted these effects on rust pustule size. Similar effects were observed with cv. Mazurka where inoculations with E. graminis produced only small necrotic flecks but did induce premature loss of chlorophyll. On this cultivar (in contrast to Zephyr) the inoculation of one leaf surface affected the development of P. hordei on the other. In comparable experiments using Zephyr, E. graminis produced smaller colonies with fewer conidiophores on leaves previously inoculated with P. hordei. These effects could be alleviated by arresting rust development with a spray containing benodanil or by floating segments of leaves inoculated with both fungi on 2% sucrose. Germination of the conidia of E. graminis, formation of appressoria and initiation of colonies were not affected by the presence of P. hordei.     

Factors affecting aphid acquisition and transmission of soybean mosaic virus

Myzus persicae transmitted soybean mosaic virus (SMV) most efficiently following 30 or 60 s acquisition probes on infected plants. There were no differences in susceptibility to SMV infection of soybean plants 1 to 12 wk old, but symptoms were more severe in plants inoculated when young than when old. Soybeans inoculated between developmental stages R3 and R6 only showed yellowish-brown blotching on one or more leaves. There were no observable differences in the time of appearance or type of symptoms shown by soybean seedlings inoculated either by sap or by aphids; infected plants became acquisition hosts for aphids 5–6 days after inoculation. There was no change in the efficiency with which M. persicae transmitted SMV from source plants up to 18 wk after inoculation. M. persicae transmitted SMV from leaves of field-grown soybeans when plants were inoculated at developmental stages V6, R2, and R3 and tested as sources 57–74 days after inoculation but not from plants inoculated at R5 and tested as sources 14 to 32 days after inoculation. M. persicae acquired SMV from soybean buds, flowers, green bean pods, and unifoliolate, trifoliolate, and senescent leaves. Middle-aged and deformed leaves were better sources of the virus than buds, unfolding and old symptomless leaves. The results are being incorporated into a computer model of SMV epidemiology.     

Effect of Leaf Age and Nodal Position on Receptivity of Rice Leaves to Infection by Pyricularia grisea

The effect of leaf age and nodal position on leaf receptivity to rice blast, caused by Pyricularia grisea , was studied by inoculating potted Rosemont plants with blast pathogen race IC-17, isolate 92T107. at different growth stages. Regressioti equations were used to describe the effect ofleaf age (degree days > 10 C) and nodal position on lesion density, the relative frequency of lesion appearance, and the lesion area distribtition. The number of sporulating lesions per cm² of inoculated leaf area was highest for leaves inoculated early in their development and progressively lower for leaves inoculated later in their development. The average lesion density for less than 1-day-old leaves at nodal position 6 was about four times the density for 10-day-old leaves of the same nodal position. Lesion density was similar for leaves of less than 1-day-old from the 4th to the 7th nodal position, but dropped sharply from the 7th to the 11 th nodal position. The flag leaf was the least susceptible of all nodal positions. The average lesion density for young leaves of about 1-day-old at nodal position 6 was about 27 times the density for young flag leaves of the same age. The incubation period increased with leaf age. but was not obviously affected by leaf nodal position. Lesion area increased linearly with time, but was not affected by leaf age and nodal position. The results are of use for epidemiological investigations which rely on detailed quantitative leaf susceptibility data for accurate blast forecasts.     

Characterization and Comparison of Clavibacter michiganensis subsp. nebraskensis Strains Recovered from Epiphytic and Symptomatic Infections of Maize in Iowa

Clavibacter michiganensis subsp. nebraskensis (Cmn), the causal organism of Goss’s wilt and leaf blight of maize, can be detected in the phyllosphere of its host prior to disease development. We compared the morphology and pathogenicity of 37 putative isolates of Cmn recovered from asymptomatic and symptomatic maize leaves. Thirty-three of the isolates produced mucoid orange colonies, irrespective of the source of isolation and all but four of these isolates were pathogenic on maize. The remaining 4 isolates recovered from asymptomatic leaves had large fluidal yellow colonies, and were non-pathogenic on maize. Isolates varied in their aggressiveness on a susceptible hybrid of maize but no significant differences in aggressiveness were detected between epiphytic isolates and those recovered from diseased maize tissues. The genomics of Cmn is poorly understood; therefore as a first step to determining what genes may play a role in virulence, we compared 33 putative virulence gene sequences from 6 pathogenic and a non-pathogenic isolate recovered from the phyllosphere. Sequence polymorphisms were detected in 5 genes, cellulase A, two endoglucanases, xylanase B and a pectate lyase but there was no relationship with pathogenicity. Further research is needed to determine what genes play a role in virulence of Cmn. Our data show however, that the virulence factors in Cmn likely differ from those reported for the closely related subspecies michiganensis and sepedonicus.     

Differentiation of Two New York Isolates of Pratylenchus penetrans Based on Their Reaction on Potato

The behavior of two isolates of Pratylenchus penetrans on six potato clones was assessed to test the hypothesis that these nematode isolates from New York were different. Four potato cultivars (Superior, Russet Burbank, Butte, and Hudson) and two breeding lines (NY85 and L118-2) were inoculated with nematode isolates designated Cornell (CR) and Long Island (LI). Population increase and egression of nematodes from roots were used to distinguish resistance and susceptibility of the potato clones. Based on numbers of eggs, juveniles, and adults in their roots 30 days after inoculation, potato clones Butte, Hudson, and L118-2 were designated resistant to the CR isolate and susceptible to the LI isolate. More eggs were found in the roots of all plants inoculated with the LI isolate than with the CR isolate. The clones NY85 and L118-2 were inoculated with the CR and LI isolates in a 2 x 2 factorial experiment to assess differences in nematode egression. Egression was measured, beginning 3 days after inoculation, for 12 days. The rates of egression were similar for the four treatments and fit linear regression models, but differences were detected in numbers of egressed nematodes. More nematodes of the CR isolate than the LI isolate egressed from L118-2. Differences in egression of females was particularly significant and can be used as an alternative or supplement to reproduction tests to assess resistance in potato to P. penetrans and to distinguish variation in virulence.     

Inhibitory effect of fucoidan from brown alga Fucus evanescens on the spread of infection induced by tobacco mosaic virus in tobacco leaves of two cultivars

The effect of fucoidan from the brown alga Fucus evanescens on the spread of infection induced by tobacco mosaic virus (TMV) was investigated in the leaves of tobacco (Nicotiana tabacum L.) of two cultivars (Ksanti-nk and Samsun). In the leaves of cv. Ksanti-nk inoculated with a mixture of TMV preparation (2 μg/ml) and fucoidan (1 mg/ml), the number of local necrotic lesions induced by the virus decreased by more than 90% as compared with the leaves inoculated with the virus alone. In tobacco leaves of cv. Samsun, virulence and the concentration of the virus 3 days after inoculation with the same mixture of TMV and fucoidan were by 62 and 66%, respectively, lower than in the leaves inoculated with TMV alone. As the infection spread, the inhibitory effect of fucoidan decreased. When the leaves were treated with fucoidan before and after the inoculation with TMV, its antiviral activity was less pronounced than when a mixture of the virus and the polysaccharide was used as inoculum. Electron microscopic investigation of TMV mixed with fucoidan often showed agglutinated virions. The highest virulence of the mixture (TMV preparation, 12 μg/ml, plus fucoidan, 1 mg/ml) was observed upon its twofold dilution, and after that it decreased. It was concluded that, when the leaves were inoculated with the mixture of TMV and fucoidan, the latter affected not only the plant but the virus as well. Treatment of tobacco leaves, cv. Ksanti-nk, with actinomycin D (10 μg/ml) 24 h before the inoculation with TMV almost completely suppressed the effect of fucoidan, indicating that fucoidan acted at a gene level.     

The gall miteAceria cladophthirus. I. Life-cycle,survival outside the gall and symptoms' expression on susceptible or resistantSolanum dulcamara plants

The gall miteAceria cladophthirus (Nalepa) is able to survice outside its gall on detached leaves ofSolanum dulcamara L. kept under non-aseptic in-vitro conditions. The survival rate of the females on susceptible leaves is about 90% after 1 day and 85% for the following days. In contrast, on resistant leaves, less than 40$ survive after 1 day while necrotic local lesions develop and later the mortality increases severely. However, the mite only completes its life-cycle on susceptible leaves. The life-cycle forA. cladophthirus takes about 12 days: six days for egg incubation and six days for two instars growth. Its life-history is simple, without alternating females specialised for hibernation; arrhenotokous parthenogenesis occurs in experimental conditions. Eggs do not play any role in gall formation. Immature stages induce gall symptoms but are less efficient than females. Mite feeding only induces complete gall symptoms on the less differentiated leaves of susceptible shoots. On susceptible detached leaves, gall symptoms are similar but weaker and their intensity decreases with increasing leaf age; fully expanded leaves remain free of symptoms.On resistant plants, mite feeding induces a hypersensitive response: necrotic local lesions, about 350 m in diameter, appear both on shoots and on detached leaves. However, young leaves develop smaller lesions than old ones. Females induce larger lesions than first larvae. Once necrosis formation is initiated, it proceeds to completion regardless of the duration of mite feeding.     

Seedling Blights of Cyperaceae Weeds Caused by Curvularia tuberculata and C. oryzae

Two Curvularia tuberculata isolates (93-020 and 93-022) and one C. oryzae isolate (93-061) were obtained from diseased Cyperus difformis , C. iria , and Fimbristylis miliacea , respectively, in the Philippines in 1993. When inoculated onto their hosts at 1 ×10 8 conidia m -2 , and provided with 24-h dew at 28°C, a severe and rapid blighting of the leaves caused the weed seedlings to die within two weeks. The three Curvularia isolates were cross-pathogenic to C. difformis , C. iria , F. miliacea , and C. rotundus but exhibited varying degrees of virulence depending on the weed species being attacked. C. tuberculata isolates 93-020 and 93-022 effectively controlled C. difformis and C. iria but were less effective on F. miliacea.

C. oryzae was equally virulent on C. difformis , C. iria , and F. miliacea . C. rotundus was not killed by any of the three isolates and exhibited only minor flecking and leaf tip die-back. Sporulation of the three Curvularia isolates on necrotic leaf tissues occurred on the susceptible ( C. difformis , C. iria , and F.miliacea ) but not on the resistant ( C. rotundus ) weed hosts.     

Inheritance of powdery mildew (Erysiphe polygoni DC) resistance in mungbean (Vigna radiata L. Wilczek)

Detached mungbean (Vigna radiata L.Wilczek) leaves were inoculated with a conidial suspension of a local isolate (TI-1) of the powdery mildew pathogen (Erysiphe polygoni DC) under controlled environment conditions. Based on the latent period and severity of the infection, a rating scale of 0–5 was used to classify the host pathogen interactions. Reactions 0, 1 and 2 were considered resistant and referred to as R0, R1 and R2 while 3, 4 and 5 were classified as susceptible (S). RUM lines (resistant to powdery mildew) and their derivatives are crossed with several susceptible (reaction types 3–5) genotypes and the inheritance of the resistance was studied in the F₁, F₂ and F₃ generations. The results showed that powdery mildew resistance in mungbean is governed by two dominant genes designated as Pm-1 and Pm-2. When both Pm-1 and Pm-2 were present, an R0 reaction was observed after inoculation with TI-1. The resistant reaction was R1 when only Pm-1 was present and R2 in the presence of Pm-2. In the absence of both Pm-1 and Pm-2, susceptible reactions 3, 4 and 5 were observed.     

Selection, biological properties and fitness of resistance-breaking strains of Tomato spotted wilt virus in pepper

In glasshouse tests, infective sap from plants infected with 17 different isolates of Tomato spotted wilt virus (TSWV) from four Australian states was inoculated to three Capsicum chinense accessions (PI 152225, PI 159236 and C00943) carrying single genes that confer hypersensitive resistance to TSWV. The normal response to inoculation was development of necrotic (hypersensitive) local lesions in inoculated leaves without systemic invasion, but 3/1386 infected plants also developed systemic susceptible reactions in addition to hypersensitive ones. Similarly when two isolates were inoculated to C. chinense backcross progeny plants, 1/72 developed systemic susceptible reactions in addition to localised hypersensitive ones. Using cultures from the four plants with susceptible reactions and following three to five further cycles of serial subculture in TSWV‐resistant C. chinense plants, four isolates were obtained that gave systemic susceptible type reactions in the three TSWV‐resistant accessions, and in TSWV‐resistant cultivated pepper (C. annuum). When three of these isolates were inoculated to tomato (Lycopersicon esculentum) breeding lines with single gene resistance to TSWV, resistance was not overcome. Similarly, none of the four isolates overcame partial resistance to TSWV in Lactuca virosa. When TSWV isolates were inoculated to tomato breeding lines carrying partial resistance from L. chilense, systemic infection developed which was sometimes followed by ‘recovery’. After four successive cycles of serial passage in susceptible cultivated pepper of a mixed culture of a resistance‐breaking isolate with the non resistance‐breaking isolate from which it came, the resistance‐breaking isolate remained competitive as both were still found. However, when the same resistance‐ breaking isolate was cultured alone, evidence of partial reversion to wild‐type behaviour was eventually obtained after five but not four cycles of long term serial subculture in susceptible pepper, as by then the culture had become a mixture of both types of strain. This work suggests that resistance‐breaking strains of TSWV that overcome single gene hypersensitive resistance in pepper are relatively stable. The findings have important implications for situations where resistant pepper cultivars are deployed widely in the field without taking other control measures as part of an integrated TSWV management strategy.     

Growth of Pseudomonas phaseolicola in susceptible and in resistant bean plants

Race 1 of Pseudomonas phaseolicola introduced into leaves of susceptible Canadian Wonder bean plants multiplied logarithmically for 3–5 days, reaching final populations about 10⁵–10⁶ times the original. In resistant Red Mexican, Race 1 multiplied less rapidly to give final populations about 10²–10³ times the original. Race 2 behaved in susceptible Red Mexican as did Race 1 in Canadian Wonder. Macroscopic symptoms appeared in leaves when bacterial numbers reached their maxima. When introduced into the cotyledonary node Race 1 moved more rapidly upwards than downwards, and more rapidly and farther in Canadian Wonder than in Red Mexican. But even in Canadian Wonder the bacterium appeared only sporadically above the node of the first compound leaf. It could be isolated only rarely from chlorotic haloes around necrotic areas in leaves, or from chlorotic leaves not carrying lesions. Fewer lesions developed and the bacteria multiplied less in older than in younger leaves. Addition of glucose and casein hydrolysate to inocula of Race 1, separately or together, had little effect on growth in Canadian Wonder or Red Mexican, and the bacterium grew equally well in extracts of susceptible and of resistant plants. Preinoculation of leaves with an avirulent race reduced the number of lesions caused by a virulent race inoculated later, and also reduced growth of this race in leaves of a susceptible variety.     

Partial Characterization of a Gene for a 31 kD Protein of Pseudomonas syringae pv. syringae Apparently Involved in Pathogenicity

P. syringae pv. syringae strain R32 causes the bacterial brown spot disease on bush beans. A 31 kD protein was detected which is involved in the pathogenic response. Monospecific antibodies directed against this 31 kD-protein were used to screen a protein expression gene bank made from the wild type strain R32. A 0.8 kb DNA insert of a clone which gave a positive reaction with the monospecific antibodies was used in hybrizations to clone a larger chromosomal fragment. A km-cassette (kmresistance) was integrated into this chromosomal DNA-fragment preventing the expression of the 31 kD-protein. This construct was integrated into the chromosomal of the wild type strain R32 viahomologous recombination resulting in the 31 kD-protein deficient mutant LMI. Biotests with the host plant (bean) and with tobacco leaves showed no symptoms or hypersensitive reaction (HR) when the mutant LMI was inoculated. However, atypical chlorotic and necrotic lesions compared to the wild type strain R32 were found on tobacco leaves when the mutant LMI was incubated for more than 2 weeks. Complementation of the LMI mutant with a plasmid harbouring the corresponding wild type R32 DNA fragment resulted in an isolate (LMIC) which showed a partial restoration of the HR on tobacco but no brown spot disease symptoms on bush beans. The 31 kD-protein could be detected serologically in LMIC.     

The Potential of Stagonospora sp. as a Mycoherbicide for Field Bindweed

Field bindweed ( Convolvulus arvensis L.) is one of the 12 most important weeds worldwide. Stagonospora sp. Isolate LA39 was isolated from diseased field bindweed plants collected in Europe. No crop tested was susceptible to the fungus, but disease symptoms were observed on other Convolvulaceae species. On field bindweed, the fungus induces disease symptoms (i.e. lesions) mainly on leaves and less severely on stems. The application of spores in an oil emulsion (10% oil in water) enhanced the disease on field bindweed plants compared with spores suspended in a 0.1% aqueous solution of the surfactant agent, Tween 80. The necrotic leaf area of inoculated plants increased as the length of exposure to 100% relative humidity (RH) and the spore density applied increased. Severe disease developed on plants inoculated with 1 107 spores/ml in oil emulsion, even in the absence of exposure to 100% RH. A delay of exposure to 100% RH (up to 8 h) did not have a significant eVect on disease severity. Field bindweed was susceptible to the fungus at all growth stages tested, but older plants were more susceptible than younger ones. It was concluded that isolate LA39 has potential as a biocontrol agent of field bindweed, especially when applied in an oil emulsion. The oil emulsion maintains the aggressiveness of the pathogen during a dew-free period and provides a favourable microenvironment during the infection process.     

The phytotoxin coronatine from Pseudomonas syringae pv. tomato DC3000 functions as a virulence factor and influences defence pathways in edible brassicas

The phytotoxin coronatine (COR) contributes to the virulence of Pseudomonas syringae pv. tomato ( Pst ) strain DC3000 on Arabidopsis thaliana and tomato. However, little is known regarding the role of COR in the virulence of DC3000 on cultivated Brassica spp. In this study, the role of COR and its precursors, coronafacic acid (CFA) and coronamic acid (CMA), were examined in the virulence of Pst DC3000 on collard and turnip, two important edible brassicas. Pst DC3000 and three well-defined COR⁻ biosynthetic mutants of DC3000 exhibited substantial differences in the timing and phenotype of disease lesions on collard and turnip. When examined 3 days post-inoculation (dpi), collard inoculated with DC3000 exhibited visible anthocyanin production and lesions were chlorotic and water-soaked. On turnip, chlorotic and necrotic lesions were evident on DC3000-inoculated leaves 5 dpi. The bacterial population dynamics on plants inoculated with DC3000 and the COR⁻ mutants indicated that COR was essential for DC3000 to maintain high populations in turnip, but not collard. Real-time quantitative PCR revealed that the jasmonic acid pathway responsive genes, LOX2 and CORI1 , were expressed in both hosts inoculated with Pst DC3000. PR1 , a marker associated with the salicylic acid pathway, was expressed in collard and turnip inoculated with the CFA⁻ CMA⁻ mutant DB29, but not DC3000. Further comparison of PR1 and LOX2 expression indicated that CFA plays a subtle role in modulating defence in turnip. This is the first study to investigate the role of COR in the interaction of Pst DC3000 and cultivated brassicas using genetically and biochemically defined COR⁻ mutants.     

BALB/c mice infected with antimony treatment refractory isolate of Leishmania braziliensis present severe lesions due to IL-4 production

Background

Leishmania braziliensis is the main causative agent of cutaneous leishmaniasis in Brazil. Protection against infection is related to development of Th1 responses, but the mechanisms that mediate susceptibility are still poorly understood. Murine models have been the most important tools in understanding the immunopathogenesis of L. major infection and have shown that Th2 responses favor parasite survival. In contrast, L. braziliensis–infected mice develop strong Th1 responses and easily resolve the infection, thus making the study of factors affecting susceptibility to this parasite difficult.

Methodology/Principal Findings

Here, we describe an experimental model for the evaluation of the mechanisms mediating susceptibility to L. braziliensis infection. BALB/c mice were inoculated with stationary phase promastigotes of L. braziliensis, isolates LTCP393(R) and LTCP15171(S), which are resistant and susceptible to antimony and nitric oxide (NO), respectively. Mice inoculated with LTCP393(R) presented larger lesions that healed more slowly and contained higher parasite loads than lesions caused by LTCP15171(S). Inflammatory infiltrates in the lesions and production of IFN-γ, TNF-α, IL-10 and TGF-β were similar in mice inoculated with either isolate, indicating that these factors did not contribute to the different disease manifestations observed. In contrast, IL-4 production was strongly increased in LTCP393(R)-inoculated animals and also arginase I (Arg I) expression. Moreover, anti-IL-4 monoclonal antibody (mAb) treatment resulted in decreased lesion thickness and parasite burden in animals inoculated with LTCP393(R), but not in those inoculated with LTCP15171(S).

Conclusion/Significance

We conclude that the ability of L. braziliensis isolates to induce Th2 responses affects the susceptibility to infection with these isolates and contributes to the increased virulence and severity of disease associated with them. Since these data reflect what happens in human infection, this model could be useful to study the pathogenesis of the L. braziliensis infection, as well as to design new strategies of therapeutic intervention.     

Accelerated leaf senescence takes part in enhanced resistance in cucumber mosaic virus inoculated pepper leaves

Altered photosynthetic reactions in cucumber mosaic virus (CMV) inoculated leaves of virus resistant lines L113 and L57 and susceptible pepper (Capsicum annuum L.) plants cv. Albena grown in controlled environment and in the field were investigated. The CMV inoculated leaves of virus resistant lines developed different symptoms—necrotic local lesions on L113 and chlorotic spots on L57 while the same leaves of susceptible cv. Albena were symptomless. The changes in Photosystem II (PSII) and PSI electron transport were evaluated by chlorophyll fluorescence, and far-red (FR) light induced leaf absorbance A _810–860. CMV infection caused a decrease in maximal PSII quantum yield, F _v/F _m, in susceptible leaves. Increased non-photochemical fluorescence quenching in CMV-inoculated leaves of both resistant lines were observed. In CMV-inoculated leaves of all tested plants FR light induced P700 oxidation was decreased. In the present study, the viral-infected pepper plants grown in controlled environment to avoid the effects of abiotic factors were used as model system that allow us to investigate the differences in leaf senescence in CMV-inoculated leaves of susceptible and resistant pepper lines expressing different symptoms. Earlier leaf falls of inoculated leaves as a result of accelerated leaf senescence is important for building successful secondary virus resistance strategy following fast responses such as hypersensitive reaction.